Replacement of a single cysteine in the fourth transmembrane region of zebrafish pannexin 1 alters hemichannel gating behavior.

Pannexin1 (Panx1) is a novel candidate for an electrical synapse protein in the retina. At present Panx1 is considered to function as a hemichannel. Since information about the gating properties of Panx1 channels to date rely on blocker pharmacology, we have begun to establish a structural context of channel function starting with site directed mutagenesis of cysteine residues in transmembrane domains of Panx1. Dye uptake and whole cell voltage clamp recordings of transfected N2a cells demonstrate that zfPanx1 forms voltage activated hemichannels with a large unitary conductance in vitro. The function of this channel was significantly reduced following mutation of a single cysteine residue (C282W) in the fourth transmembrane region. This result suggests a role of this domain in gating of the Panx1 hemichannel.